Introduction

Neural crest (NC)-like stem/progenitor cells provide an attractive cell source for regenerative medicine because of their multipotent property and ease of isolation from adult tissue. Although human umbilical cord blood (HUCB) is known to be a rich source of stem cells, the presence of the NC-like stem/progenitor cells in HUCB remains to be elucidated. In this study, we have isolated NC-like progenitor cells using an antibody to p75 neurotrophin receptor (p75NTR) and examined their phenotype and stem cell function in vitro.

Methods

To confirm whether p75NTR⁺ NC-derived cells are present in cord blood, flow cytometric analysis of cord blood derived from P0-Cre/Floxed-EGFP reporter mouse embryos was performed. Freshly isolated HUCB mononuclear cells was subjected to flow cytometry to detect p75NTR⁺ cells and determined their immunophenotype. HUCB p75NTR⁺ cells were then collected by immunomagnetic separation and their immunophenotype, clonogenic potential, gene expression profile, and multilineage differentiation potential were examined.

Results

NC-derived EGFP⁺ cells co-expressing p75NTR was detected in cord blood of P0-Cre/Floxed-EGFP reporter mice. We found that freshly isolated HUCB mononuclear cells contained 0.23% of p75NTR⁺ cells. Isolated p75NTR⁺ cells from HUCB efficiently formed neurospheres and could differentiate into neuronal and glial cell lineages. The p75NTR⁺ cells expressed a set of NC-associated genes and undifferentiated neural cell marker genes before and after the culture.

Conclusions

These findings revealed that HUCB contained the p75NTR⁺ NC-like progenitor cell population which have the self-renewal capacity and the potential to differentiate into both neuronal and glial cell lineages.

中文翻译：

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人脐带血中神经嵴样祖细胞的分离与鉴定

介绍

神经嵴 (NC) 样干/祖细胞为再生医学提供了有吸引力的细胞来源，因为它们具有多能特性并且易于从成人组织中分离。尽管已知人脐带血 (HUCB) 是丰富的干细胞来源，但 HUCB 中是否存在类似 NC 的干/祖细胞仍有待阐明。在这项研究中，我们使用 p75 神经营养因子受体 (p75NTR) 的抗体分离了 NC 样祖细胞，并在体外检测了它们的表型和干细胞功能。

方法

为了确认 p75NTR ⁺ NC 衍生细胞是否存在于脐带血中，对源自 P0-Cre/Floxed-EGFP 报告小鼠胚胎的脐带血进行了流式细胞术分析。对新鲜分离的HUCB单核细胞进行流式细胞术以检测p75NTR ⁺细胞并确定它们的免疫表型。然后通过免疫磁性分离收集HUCB p75NTR ⁺细胞，并检查它们的免疫表型、克隆形成潜力、基因表达谱和多向分化潜力。

结果

在 P0-Cre/Floxed-EGFP 报告小鼠的脐带血中检测到NC 衍生的 EGFP ^{+细胞共表达 p75NTR。}我们发现新鲜分离的 HUCB 单核细胞含有 0.23% 的 p75NTR ⁺细胞。从 HUCB分离的 p75NTR ⁺细胞有效地形成了神经球，并且可以分化成神经元和神经胶质细胞谱系。p75NTR ⁺细胞在培养前后表达了一组 NC 相关基因和未分化的神经细胞标记基因。

结论

这些发现表明，HUCB 含有 p75NTR ⁺ NC 样祖细胞群，其具有自我更新能力和分化成神经元和神经胶质细胞谱系的潜力。